Expression of a lipoxygenase encoding gene (BoLOX1) during postharvest senescence of broccoli

Lipoxygenases (LOX) belong to a large family of plant enzymes that catalyze the hydroperoxidation of polyunsaturated fatty acids. Most of them are expressed during senescence and contribute to membrane deterioration and biosynthesis of jasmonic acid, a known senescence enhancer. In this work, we cloned a fragment of a gene encoding a LOX from broccoli (BoLOX1). The analysis of the sequence revealed that BoLOX1 is closely related to other LOX from higher plants. Furthermore, we analyzed the expression of BoLOX1 and detected a larger increase during postharvest senescence. A slight increase of total lipoxygenase activity was also found during senescence. In other sets of experiments, broccoli heads were treated with plant hormones, such as cytokinin and ethylene, as a way to assess the effect of such compounds on the expression of BoLOX1. Cytokinin treatment delayed the increase of BoLOX1 expression and lipoxygenase activity whereas ethylene accelerated both processes. Also, several postharvest treatments were applied in order to delay senescence in broccoli florets and to evaluate their effects on BoLOX1 expression. Samples treated with modified atmosphere, hot air, UV-C or white light showed a delay in chlorophyll degradation and degreening. In most cases, the treatments also delayed the increase of BoLOX1 expression, reaffirming the relationship between the expression of this gene and senescence. However, treatments like modified atmospheres and visible light markedly increased lipoxygenase activity, which suggests a lack of correlation between BoLOX1 expression and lipoxygenase activity.     

烤烟立体育苗适宜人工光源筛选研究

为筛选出适宜烤烟立体育苗生长的人工光源,在全黑实验室条件下对4种人工光源进行了光照环境指标的测试,分析比较了距灯管距离为20、30、40 cm时光照的强度和均匀性;设置4个不同光照条件下烤烟幼苗栽培试验区,对成苗生理生态指标进行了数据统计和分析。结果表明：（1）4种人工光源平均光照强度从大到小排序依次为红蓝LED>全光谱荧光灯>暖白LED>三基色荧光灯;（2）距灯管距离30 cm的光照分布均匀性最好,其次为20、40 cm。（3）栽培试验区采用红蓝LED灯补光,光照强度设置为150 μmol/(m²·s)时,成苗品质最好。研究结果为烤烟立体育苗人工补光配置方案提供理论依据。     

Expression of Stay-Green encoding gene (BoSGR) during postharvest senescence of broccoli

Degreening caused by chlorophyll degradation is the most important feature that determines postharvest loss of quality in broccoli. Chlorophyll molecules are assembled to several thylakoid proteins, from which chlorophylls must be released in order to be catabolized. Stay-Green (SGR), a chloroplast-located protein, specifically interacts with light harvesting complex subunits helping toward their destabilization and to the release of chlorophylls. In this work, a fragment of a gene encoding a SGR from broccoli (BoSGR) was cloned. The expression of BoSGR was analyzed and detected an important increment during postharvest senescence, simultaneously with chlorophyll degradation. In order to analyze the effect of different growth regulators, different groups of broccoli heads were treated with cytokinins, ethylene and 1-MCP. Cytokinins and 1-MCP delayed the increment of BoSGR expression whereas ethylene accelerated the process. In addition, several postharvest treatments that delay degreening in broccoli florets were applied to evaluate their effects on BoSGR expression. Samples treated with modified atmosphere, hot air, UV-C or white lights showed a delay in chlorophyll degradation and degreening. In most cases, the treatments also delayed the increment of BoSGR expression during senescence, reaffirming the relationship between the expression of this gene and chlorophyll degradation.     

马铃薯组培苗时期不同LED光源处理对温室移栽后植株生长和微型薯结薯的影响

不同LED光源下培养的马铃薯组培苗在形态特征上差异明显,为探索这种差异是否会对其移栽后产生进一步影响,试验研究了100%红光(R)、100%蓝光(B)、100%绿光(G)、75%红光+25%蓝光(RB)、45%红光+35%蓝光+20%绿光(RBG)和100%白光(CK)6种光谱下培养4周的马铃薯组培苗温室移栽后植株的生长和结薯情况。试验结果表明,B处理的马铃薯组培苗移栽后缓苗期叶绿素荧光参数Fv/Fm和qP、结薯前期SPAD值及结薯期各器官生物量均始终高于CK和其他处理,且该处理下最大薯的干鲜重和有效微型薯占比分别较CK提高45.66%、45.09%和10.93%。R处理的组培苗移栽后结薯期地上部生物量显著高于CK,单株结薯数目提高29.60%,但有效微型薯占比较CK降低63.97%。G处理的组培苗移栽后生长和结薯指标均表现最差。组合光谱RB和RBG处理的组培苗移栽后的生长指标和微型薯产量及产量构成指标均与CK无显著差异。因此,单色红、蓝、绿光谱处理对马铃薯组培苗的影响会持续至其移栽后植株幼苗生长,甚至结薯期,组合光谱RB和RBG则不会。在马铃薯组培苗生产上,单色蓝光和单色红光作为照明光源具有潜在的应用价值,组合光谱RB和RBG可作为白色光源的替代光源。     

Application of low intensity light pulses to delay postharvest senescence of Ocimum basilicum leaves

Fresh basil (Ocimum basilicum L.) is a highly perishable leafy green vegetable with a storage life of 4–5 d at room temperature. Exposure of basil leaves to temperatures below 12 °C during storage results in chilling injury; therefore, refrigeration cannot be used to extend postharvest life of basil. Typically, leafy vegetables are stored in darkness or extremely low irradiance. Darkness is known to induce senescence, and the initial phase of senescence is reversible by exposure to light. In this work, we studied the effects of low-intensity white light pulses at room temperature on postharvest senescence of basil leaves. Daily exposure for 2 h to 30–37 μmol m⁻² s⁻¹ of light was effective to delay postharvest senescence of basil leaves. Chlorophyll and protein levels decreased, ammonium accumulated and leaves developed visual symptoms of deterioration (darkening) during storage in darkness. Light pulses reduced the intensity of these senescence symptoms. The photosynthesis light compensation point of basil leaves was 50 μmol m⁻² s⁻¹ i.e., higher than the intensity used in this study, and the effect of treatment with red light was the same as with white light, while far red light was ineffective. Light pulses exerted a local effect on chlorophyll loss, but the effect on protein degradation was systemic (i.e., spreading beyond the illuminated parts of the leaf blade). The results of this study indicate that daily treatment for 2 h with low intensity light (30–37 μmol m⁻² s⁻¹ every day) during storage at 20 °C is an effective treatment to delay postharvest senescence of basil leaves. The delay of postharvest senescence by low intensity light pulses seems to be mediated by phytochromes, and it is systemic for protein, and partially systemic for chlorophyll degradation.     

LED effect on free radical-scavenging activity of barley leaf

Barley leaf grown under light-emitting diode (LED) light was prepared and investigated for its free radical scavenging activity. LED light such as red, far-red, blue, blue-red, green, yellow, UVA, and white and also fluorescent light and dark conditions were also used in this study. Length and weight of barley leaves were measured and the Hunter color value, DPPH radical scavenging activity, and ABT radical-scavenging activity of barley leaf extracts were estimated. Extraction of barley leaf was performed by using a 70% ethanol solution. Length of barley leaf grown with red and green LED was 9.9 and 12.6 cm, respectively. Hunter L* and a* values of extracts of barley leaf grown under dark conditions were higher than those of other LED light-treated samples. The DPPH radical scavenging activity of extracts of barley leaf grown with blue, UVA, and under dark conditions were 41.40, 36.23, and 43.57%, respectively. ABT radical scavenging-activity of extracts of barley leaf grown under LED of red, far-red, blue, blue-red, green, yellow, and white were 5.46, 6.10, 6.00, 5.93, 5.74, 6.32, and 5.94 μmol TE g⁻¹, respectively.     

LED不同光质对菘蓝愈伤组织诱导及再生的影响

[目的]为研究不同光质对菘蓝愈伤组织诱导及再生的影响,[方法]以定蓝1号菘蓝种子为材料,设置6个LED光照处理,分别为：荧光灯(CK),100R,100B,80R/20B,70R/30B,50R/50B,比较不同光质对菘蓝愈伤组织诱导、再生及生根情况,同时测定再生植株叶绿素含量及可溶性糖含量。[结果]结果表明：红蓝混合光处理较单一红蓝光有明显优势,70R/30B处理下,菘蓝愈伤组织愈伤生长状态最好,表现为颜色绿,质地致密,分化率最高为68.75%,再生植株生根率最高为92.86%,生根状态表现为生根早,根粗壮,根长等特征,均显著高于对照。70R /30B 处理下,叶绿素a、总叶绿素含量及可溶性糖含量均达到最大值。[结论]综上,红蓝混合光处理有利于菘蓝愈伤组织的诱导分化,再生植株的形态建成,叶绿素合成、可溶性糖积累,其中以70R/30B为最适光源。     

UV-C treatment delays postharvest senescence in broccoli florets

Central broccoli heads (cv. de Cicco) were harvested and treated with UV-C light (4, 7, 10, or 14 kJ m⁻²). All treatments delayed yellowing and chlorophyll degradation at 20 °C but the irradiation dose of 10 kJ m⁻² allowed retaining the highest chlorophyll content yet had lower amounts of pheophytins than every treatment other than 7 kJ m⁻². This dose was selected to analyze the effect of UV-C on postharvest broccoli senescence at 20 °C. The UV-C treatment delayed yellowing, chlorophyll a and b degradation, and also the increase in pheophytins during storage. The activity of chlorophyll peroxidase and chlorophyllase was lower in UV-C treated broccoli. Instead, Mg-dechelatase activity increased immediately after the treatment, but after 4 and 6 d this activity was lower in UV-C treated florets than in controls. Treated broccoli also displayed lower respiration rate, total phenols and flavonoids, along with higher antioxidant capacity. The results suggest that UV-C treatments could be a useful non-chemical method to delay chlorophyll degradation, reduce tissue damage and disruption, and maintain antioxidant capacity in broccoli.     

Effects of postharvest ethanol vapor treatment on activities and gene expression of chlorophyll catabolic enzymes in broccoli florets

Postharvest yellowing of broccoli (Brassica oleracea L. Italica Group) is an important indicator of quality deterioration and occurs with chlorophyll (Chl) degradation. Postharvest ethanol vapor treatment could delay the yellowing of broccoli florets, through the suppression of Chl degradation. As the first step in identifying the mechanism of this delay, the effects of postharvest ethanol vapor treatment on activities and gene expression of the Chl catabolic enzymes in broccoli were determined. Broccoli branchlets were placed in a perforated polyethylene bag with or without (control) an ethanol pad and stored at 20 °C in darkness. The Chl contents of the control broccoli florets decreased remarkably after three days in storage, whereas the contents of the ethanol-treated broccoli showed no significant changes except at day one. Changes in chlorophyllase activity in the ethanol-treated broccoli were similar to those of the control until three days in storage, but then the activity tended to decease. Mg-dechelatase and Chl-degrading peroxidase activities, which increased greatly with senescence in the controls, remained unchanged in the ethanol-treated broccoli. BoCLH1 expression showed changes almost similar to those of chlorophyllase activity. BoPAO expression in the control broccoli increased greatly at day one in storage and was maintained at a high level until three days. In contrast, this expression in ethanol-treated broccoli was suppressed until two days. BoRCCR expression in the control broccoli increased until two days in storage, while the expression in the ethanol-treated broccoli showed no change during storage. These results show that postharvest ethanol vapor treatment suppressed the activities and gene expression of Chl catabolic enzymes, resulting in delayed yellowing of broccoli florets.     

光质对黄瓜幼苗绿色叶片叶绿素荧光的影响

以北方白黄瓜幼苗为试材,用蓝、绿、红发光二级管(LED)单色光进行单色光和混合光照射诱导处理,分析了光质对Fp685、Fp735、Fp685/ Fp735、F685、F735、 F685/ F735和Y（II）的影响。3种LED单色光源（蓝、绿、红）照射诱导下,随着光强的增加白黄瓜叶片叶绿素荧光曲线OJIP中P点呈现快速上升趋势。其中,Fp685主要表现为蓝光>绿光>红光;而Fp735均表现为绿光下高,红蓝光下低;Fp685/ Fp735荧光比值均表现为蓝光最高,绿光和红光下较低。混合光得到的荧光曲线的P点出现早于单色光照射,且P混合>P单色光加和。混合光荧光值计算得到的F685/F735比值始终低于单色光荧光加和值计算得到的F685/F735比值。各个单色光质下的PSⅡ实际光化学量子产量表现为：Y（II）绿光> Y（II）红光> Y（II）蓝光。     

Chlorophyll fluorescence for non-destructive measurement of flavonoids in broccoli

Chlorophyll fluorescence (ChlF) by excitation with two radiation beams of different wavelengths can be used for non-destructive quantitation of epidermal flavonoids in leaves. The method was tested on marketable broccoli heads, but including red light (685 nm) and green light (530 nm) in addition to ultraviolet (UV) radiation (382 nm) and blue light (450 nm) for excitation of ChlF. As a reference, the content of flavonoids was measured with high performance liquid chromatography. To induce flavonoids, postharvest treatment during 12 days with various combinations of visible and UV radiation was tried, but the achieved changes in the flavonoid levels were not statistically significant. Instead a large natural variation of flavonoid content among the broccoli heads was used to correlate it with the fluorescence data. It was possible to estimate the content of flavonoids in flower buds by means of ChlF: relative epidermal absorbance of blue light was well correlated with flavonoid content (r = 0.69, p < 0.001). For quercetin alone the correlation was higher (r = 0.77, p < 0.001). On the other hand, relative epidermal absorbance of UV radiation or green light had a much weaker correlation with flavonoid content (r ≤ 0.40, p < 0.05). Within the broccoli heads, flower buds had 15 times higher levels of flavonoids than the floret stalks, which in turn had five times higher contents than the main stem. Repeated ChlF measurements on individual broccoli heads during cold storage could monitor small but significant changes in flower buds, possibly indicating a breakdown of constituents absorbing UV and green light.     

Pheophytinase activity and gene expression of chlorophyll-degrading enzymes relating to UV-B treatment in postharvest broccoli (Brassica oleracea L. Italica Group) florets

Pheophytinase (PPH) activity and gene expression of chlorophyll (Chl)-degrading enzymes relating to UV-B treatment in postharvest broccoli (Brassica oleracea L. Italica Group) florets were determined. PPH is involved in the dephytylation of Mg-free Chl a, pheophytin (Phy) a. However, in vitro chlorophyllase (Chlase, EC.3.1.1.14) also uses Phy a as a substrate to produce pheophorbide (Pheide) a by dephytylation. For an accurate determination of PPH activity, the PPH protein fraction was separated from Chlase protein by ammonium sulfate precipitation. The protein precipitated by 45-60% saturated ammonium sulfate included a little bit of Chlase activity and was suitable for PPH determination. PPH activity in broccoli florets treated with a UV-B dose of 19 kJ m⁻² was repressed for the first 2 d of storage at 15 °C, whereas it increased gradually with senescence of control broccoli florets. The expression level of BoCLH1 was reduced in broccoli florets on day 4 of storage, while BoCLH2 and BoCLH3 were up-regulated with UV-B treatment. A high BoPAO expression level was found in senescent broccoli florets, and the up-regulation of this gene was delayed by UV-B treatment. The highest expression level of BoPPH was found in the control, and its expression was clearly repressed by UV-B treatment on day 2 of storage. We suggest that the up-regulation of Chl-degrading enzyme genes could be delayed by UV-B treatment, resulting in the suppression of floret yellowing in stored broccoli.     

光质对番茄营养与风味品质的影响

以番茄为试验材料,在田间大棚设施条件下,选用白、蓝、黑、红、绿5种不同颜色的遮阳网,探讨不同光质对番茄营养与风味品质的影响。结果表明,在青熟期,黑色光质促进了总氮、蛋白氮以及非蛋白氮的积累。青熟期到转色期,白色光质的番茄有机酸总量较青熟期增加了4.57倍,蓝色和绿色光质提高了番茄的总糖和还原糖含量。从转色期到成熟期,番茄氨基酸含量大小表现为蓝色>绿色>黑色>红色>白色;黑色光质的番茄有机酸总量较转色期增加了27.8%;红色光质提高了番茄红素含量和Vc含量,降低了硝酸盐含量;白色、红色和绿色光质增加了番茄的总糖和还原糖含量。番茄主要风味物质(>30 μg/kg)为水杨酸甲酯、6-甲基-庚烯-2-酮、柠檬烯、反-2-己烯醛。红光使番茄水杨酸甲酯、6-甲基-庚烯-2-酮和己烯醛含量较白光增加了11.6%、20.2%和17.2%。在大田设施条件下,转色期到成熟期可以采用红色光质提高番茄果实营养品质和风味。     

利用胚挽救获得葡萄三倍体植株

为获得三倍体葡萄新种质,以葡萄四倍体‘醉金香’×二倍体‘秋黑’杂交所得幼果为试材,研究了幼果低温处理天数、培养基IAA浓度、胚珠处理方式以及不同光质等因素对胚发育及胚萌发的影响。结果表明,采集授粉后35 d的杂交幼果,先进行4℃低温处理,再取出胚珠接种,有利于胚发育和萌发,以低温40 d和30 d处理的胚发育率和萌发率最高,分别为91.8%和24.4%;培养基IAA浓度为0.2 mg/L时可显著提高胚萌发率;进行切喙处理的胚珠胚萌发率显著高于完整胚珠接种;发光二极管LED复合光(红∶蓝∶白=3∶1∶1)比日光色荧光灯(日光灯)更适合杂种胚萌发;用流式细胞仪检测了20个杂种后代株系,其中三倍体株系19个,后代三倍体株率为95.0%。     

光质对离体培养条件下绿色棉纤维发育及黄酮合成的影响

为探讨光质对绿色棉纤维生长发育和黄酮合成的影响,以绿色棉品种绿絮棉1号为材料进行离体培养,比较红光、黄光、蓝紫光、白光和暗处理条件下绿色棉胚珠鲜物质质量、纤维长度、纤维素含量、黄酮含量等生理和分子指标的差异。结果表明:光处理能够促进胚珠鲜物质质量的增加,但抑制了纤维的伸长。在纤维发育前期,各光处理抑制了蔗糖合成酶活性和蔗糖合成酶基因表达;纤维发育后期,白光处理提高了蔗糖合成酶活性,促进了蔗糖合成酶基因表达。红光、黄光和蓝紫光处理显著提高了β-1,3￣葡聚糖酶活性(10~20 DAC(Days after culture));红光、黄光和白光均能提高β-1,3￣葡聚糖基因表达量,而蓝光则抑制了基因的表达。白光促进了纤维素的合成,而蓝紫光抑制了纤维素的合成。在纤维发育初期(10 DAC),蓝紫光抑制了黄酮合成及其关键酶基因表达;而在发育后期(30~40 DAC),白光处理能够促进黄酮的合成及其关键酶基因表达。可见,光质对绿色棉纤维发育及黄酮合成具有一定的影响。     

Effect of spring fertilization on ecosystem services of organic wheat and clover relay intercrops

Nitrogen (N) deficiency and weed infestation are main factors limiting yield and yield stability in organic wheat. Organic fertilizers may be used to improve crop performance but off-farm input costs tend to limit profitability. Instead, forage legumes may be inserted into the crop rotation to improve the N balance and to control weed infestation. In opposition to simultaneous cropping, relay intercropping of legumes in organic winter wheat limits resource competition for the legume cover crop, without decreasing the performance of the associated wheat.The aim of this study is to evaluate the effect of spring organic fertilization on the performance of intercropped legumes and wheat, and on services provided by the legume cover.Two species of forage legumes (Trifolium pratense L. and Trifolium repens L.) were undersown in winter wheat (Triticum aestivum L. cv Lona) in five organic fields during two consecutive crop seasons. Organic fertilizer was composed of feather meal and applied on wheat at legume sowing. The cover crop was maintained after the wheat harvest and destroyed just before sowing maize.Spring organic nitrogen fertilization increased wheat biomass (+35%), nitrogen (+49%), grain yield (+40%) and protein content (+7%) whatever the intercropping treatment. At wheat harvest, red clover biomass was significantly higher than white clover one (1.4 vs. 0.7 t ha⁻¹). Nitrogen fertilization decreased forage legume above-ground biomass at wheat harvest, at approximately 0.5 t ha⁻¹ whatever the specie. No significant difference in forage legume biomass production was observed at cover killing. Nitrogen accumulation in legume above-ground tissues was significantly higher for white clover than for red clover. Both red and white clover species significantly decreased weed infestation at this date. Nitrogen fertilization significantly increased weed biomass whatever the intercropping treatment and decreased nitrogen accumulation in both clover species (−12%).We demonstrated that nitrogen fertilization increased yield of wheat intercropped with forage legume while the performance of legumes was decreased. Legume growth was modified by spring fertilization whatever the species.     

新型光源发光二极管(LED)下白掌组培苗移栽后生长状况研究

为了研究白掌组培苗在新型光源发光二极管(LED)下的生长状况,以亮叶白掌为试验植物材料,采用以性能稳定的超高亮度红光LED（主波长640 nm）和超高亮蓝光LED（主波长464 nm）为光源材料,探讨了新型光源LED对白掌试管苗移栽后生长的影响,研究适合白掌组培苗移栽后生长的光质条件。结果表明：株高最大值出现在50%R+50%B处理下。在叶长和叶幅方面,光质比为80%R+20%B时达到最大值,且与对照处理间有显著性差异;在根数和根系活力方面,80%R+20%B处理下达到最大值。在根长方面,50%R+50%B处理下达到最大值。在鲜重和干重方面,白掌组培苗在80%R+20%B处理下都达到最大值。在叶绿素含量方面,只有在50%R+50%B处理下高于对照处理,其他的处理也都低于对照处理。综上分析,80%R+20%B处理的白掌组培苗移栽后生长状况最好。